The first study of molecular prevalence and species characterization of <i>Cryptosporidium</i> in free-range chicken (<i>Gallus gallus domesticus</i>) from Brazil

Ewald, Maria Paula de Carvalho; Martins, Felippe Danyel Cardoso; Caldart, Eloiza Teles; Vieira, Fernando Emmanuel Gonçalves; Yamamura, Milton Hissashi; Sasse, João Pedro; Barros, Luiz Daniel de; Freire, Roberta Lemos; Navarro, Italmar Teodorico; Garcia, João Luis

doi:10.1590/S1984-29612017068

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Original Article • Rev. Bras. Parasitol. Vet. 26 (4) • Oct-Dec 2017 • https://doi.org/10.1590/S1984-29612017068 linkcopy

The first study of molecular prevalence and species characterization of Cryptosporidium in free-range chicken (Gallus gallus domesticus) from Brazil

Primeiro estudo de prevalência molecular e caracterização de espécies de Cryptosporidium em galinhas (Gallus gallus domesticus) colonial/caipiras do Brasil

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Keywords:
Cryptosporidiosis; zoonosis; public health; epidemiology

Farms	Cities	n	n-PCR	Sequencing result
Farms	Cities	n	+/%	C. meleagridis	C. baileyi	C. parvum	C. bovis	NAQ*
1	A	70	10/ 14.3	9				1
2	A	15	5/ 33.3	4				1
3	B	5	4/ 80		2			2
4	B	59	29/ 49.1	23	2	2		2
5	C	8	0/0
6	D	10	2/20	1	1
7	E	35	2/ 5.7	1	1
8		5	1/20					1
9		6	0/0
10		20	1/5	1
11		18	2/11.1	2
12		10	7/70		6			1
13	F	37	13/35.1	11				2
14	G	2	0/0
15	G	3	2/66.7	1				1
16	H	12	2/16.7		2
17		15	6/40	3	1			2
18		11	0/0
19	I	5	3/60	1		1	1
20	J	5	2/40				2
Total		351	91/25.9	57(62.6%)	15(16.5%)	3(3.3%)	3(3.3%)	13(14.3%)

18S rRNA	14	395	396	426	442	570
CryBrFRChi70 / 71 / 108	G	T	A	C	G	G
C. bovis AY741305.1	A	C	.	T	A	.
C. xiaoi FJ896043.1	A	.	T	T	A	A
HSP70	3	15	33	63	114	135	213
CryBrFRChi70, 71 / 108	C	T	G	T	G	C	T
C. bovis AY741306.1	T	.	.	C	A	T	C
C. xiaoi FJ896041.1	T	C	A	C	.	T	C
Actin	36	117	129	147	150	153	198	216	219	249	279	303	321	366	369	372	387	393	414	438	447	453
CryBrFRChi71 / 108	A	A	T	A	A	T	G	C	C	A	G	T	T	A	G	C	T	T	G	G	C	G
C. bovis AY741307.1	G	G	G	G	G	A	A	T	T	T	A	A	C	G	.	T	C	A	A	A	T	A
C. xiaoi GQ337964.1	.	.	G	G	.	A	A	T	T	G	A	.	C	G	A	T	C	A	A	.	T	.
	483	501	507	540	555	558	576	585	618	624	633	639	666	687	714	723	732	735	744	759	786
CryBrFRChi71 / 108	T	C	G	G	C	G	G	A	A	A	A	C	C	A	T	T	A	A	A	T	C
C. bovis AY741307.1	.	.	A	A	T	A	A	G	G	G	.	T	.	G	C	C	G	T	G	C	T
C. xiaoi GQ337964.1	C	T	.	A	T	A	A	G	G	.	G	T	T	.	C	.	G	T	G	C	T

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Colégio Brasileiro de Parasitologia Veterinária FCAV/UNESP - Departamento de Patologia Veterinária, Via de acesso Prof. Paulo Donato Castellane s/n, Zona Rural, , 14884-900 Jaboticabal - SP, Brasil, Fone: (16) 3209-7100 RAMAL 7934 - Jaboticabal - SP - Brazil
E-mail: cbpv_rbpv.fcav@unesp.br

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[TFN1] ^*
NAQ: Not sequenced due to low quality of DNA.